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Effects of fluorescent probe structure on the dynamics at cysteine‐34 within bovine serum albumin: Evidence for probe‐dependent modulation of the cybotactic region
Author(s) -
Baker Gary A.,
Pandey Siddharth,
Kane Maureen A.,
Maloney Todd D.,
Hartnett Ann M.,
Bright Frank V.
Publication year - 2001
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/1097-0282(200112)59:7<502::aid-bip1055>3.0.co;2-i
Subject(s) - bodipy , chemistry , guanidine , fluorescence , bovine serum albumin , cysteine , biophysics , denaturation (fissile materials) , biochemistry , nuclear chemistry , biology , physics , quantum mechanics , enzyme
We have prepared a series of bovine serum albumins (BSA) that have been site‐selectively labeled at cysteine‐34 with one of four different sulfhydryl‐selective boron dipyrromethene difluoride (BODIPY) fluorescent probes (BODIPY FL IA, BODIPY FL C 1 IA, BODIPY 530/550 IA, and BODIPY 493/503 MB). We determine how the choice of extrinsic probe structure dictates the recovered BSA‐BODIPY dynamics under thermal (10–80°C) and chemical (0–5 M guanidine hydrochloride) denaturation conditions. The results of these experiments show that the global protein dynamics are sensed equally by each fluorescent probe; however, the probe itself influences the local probe dynamics within the cybotactic region that surrounds cysteine‐34. Thus, it seems inappropriate to think of these extrinsic fluorescent probes as passive, nonparticipatory viewers of local protein dynamics. © 2001 John Wiley & Sons, Inc. Biopolymers 59: 502–511, 2001